A Pharmacognostical Study And Analysis And Identification Based On The DNA Barcode On The Herb Of Yi Nationality Metacordyceps Liangshanensis

In recent years, the state focus a growing attention on ethnic medicine,push forward development of ethnic medicine with Chinese characteristics positively.Yi nationality medicine which is famous for its particular treatment and diagnostic technology characteristic drug use and optimize drug therapy spread up to now. Under the appeal and support of national policy, pharmacologist’s long-term effort has made gratifying achievements on the side of laboratory investigation and clinical applications which they research In-depth field,extract and arrang the ancient literatures of Yi Nationality.The result affirms the huge potential value of Yi nationality medicine and makes Yi nationality medicine usher in the new opportunities.Metacordyceps liangshanensis, one of the herb of Yi nationality, called ’MuGuBuDi’in Yi nationality area and whose scientific name is ’LiangShanChongCao’, popularizes in the Sichun, Yunnan and Guizhou provinces and is endemic species in China especially. According to Sichuan Province Traditional Chinese Medicine Standard (2010edition) it is a composite consisting of the stroma of the fungus, Metacordyceps liangshanensis (Berk.) Sacc.(Fam. Hypocreaceae), parasitized on the larva of some species of insects (Fam. Hepialidae), and the dead caterpillar’Objective:With the scarcity of resources of Cordyceps, as the promising substitute of Cordyceps sinensis, Metacordyceps liangshanensis is paied more attention and becomes a hot area of research. However, the research of Metacordyceps liangshanensis starts too late to get in depth so that the most of the study are limited at the simple expression in the external properties and microscopic identification, and the objective evaluation in the medical quality is less. Especially when Cordyceps being pshed into market gradually in the health care, the products are made by the shattered medicinal materials or used as its power state mostly, so we proceed the experiment through the pharmacognostical study and DNA barcode technology targeting at resoving the problems of the simpleness of the inspection item and the difficulty of the identification of the shattered medicinal.Method:The precise samples of Metacordyceps liangshanensis are got through field collection, investigation and specimens. We applied the property identification and microscopic identification method to the distinguish the Metacordyceps liangshanensis and the adulterants.Then the DNA barcode is done with the parasitical fungus and hosted insects in the experiment, after extracting DNA of28species of Cordyceps samples, DNA were amplified and sequenced,so the sequences were got. Based on the DNA barcode Metacordyceps liangshanensis, its adulterants and its relative species can be identificated,and it provide exactly technical support for the further reseach of the species molecular identification and phylogenetics of Cordyceps nothing to do with the developmental stage and the specificity of the organ or tissue.Result:1、On the basis of the provincial standard and pertinent literature of Metacordyceps liangshanensis, combining with The characteristics of the medicinal materials, we center on on the characteristics of caterpillar and explore the interior hypha and added a few important characters of the Metacordyceps liangshanensis.2、The molecular biological identification of the samples was proceeded which were confirmed by the resource indetification:(1) To identify and distinguish Metacordyceps liangshanensis from its adulterants and its relative species by combining ITS, COI, Cytb barcode sequences and study the feasibility of this new method.(2) Applied the molecular biological methods, we engaged in the research of genetic relationship between Metacordyceps liangshanensis, Cordyceps sinensis, Cordyceps hawkesii Gray and Cordyceps tail based on the ITS sequence. The results tell us from the NJ tree, the Metacordyceps liangshanensis and Cordyceps sinensis has close genetic distance in the parasitical fungi while the Cordyceps hawkesii Gray and Cordyceps taii has close genetic distance in the parasitical fungi.(3) To analyze the DNA bar code of parasitical insect in the Cordyceps from the CO1and Cytb sequence and discuss the feasibility, and CO1sequence is more suitable for the DNA bar research.(4) Explore the genetic relationship and phylogenetics of the hosted insect of the Cordyceps combined with the CO1and Cytb sequence. The two phylogenetic NJ trees indicate the same genus belong. And then Cordyceps hawkesii Gray and Cordyceps taii are in parallel evolution with Metacordyceps liangshanensis. And they are the top series of the cladogram,that is the evolved latest one while the Cordyceps sinensis is the earliest.Conclusion:This study conducts detailed research focusing on the external properties and microscopic identification of Metacordyceps liangshanensis at the first step.Through the comparison with the adulterants,the result confirms the specificity of the characteristics, and proofs that the research has the general practical significance and application value.Then this study established the molecular methods in identification of Metacordyceps liangshanensis primarily, precise and simple, which made contribution to the objective assessment of quality and elevated standards for Metacordyceps liangshanensis and the identification of traditional Chinese medicine, the supervision of drug administration. But our research is still in its infancy and needs improving in some aspects. In the future, we will continue to this job for research, the experiments are pointed to be continue and the data are accumulated forwards.